FXR1 expression domain in Wilms tumor

Background/PurposeWilms tumor (WT) is the most common childhood kidney cancer globally. Our prior unbiased proteomic screen of WT disparities revealed increased expression of Fragile X-Related 1 (FXR1) in Kenyan specimens where survival is dismal. FXR1 is an RNA-binding protein that associates with poor outcomes in multiple adult cancers. The aim of this study therefore was to validate and characterize the FXR1 expression domain in WT.MethodsQuantitative FXR1 gene expression was compared between WT, adjacent, adult, and fetal kidney specimens. The cellular and subcellular expression domain of FXR1 was characterized across these tissues using immunoperoxidase staining. RNA-sequencing of FXR1 was performed from WT and other pediatric malignancies to examine its broader target potential.ResultsFXR1 was detected in all clinical WT specimens evaluated (n = 82), and as a result appeared independent of demographic, histology, or adverse event. Specific cytosolic staining was strongest in blastema, intermediate and variable in epithelia, and weakest in stroma. When present, areas of skeletal muscle differentiation stained strongly for FXR1. qPCR revealed increased FXR1 expression in WT compared to adult and adjacent kidney (p < 0.0002) but was similar to fetal kidney (p = 0.648). RNA-sequencing revealed expression of FXR1 in multiple pediatric tumors, greatest in rhabdomyosarcoma and WT.ConclusionsFXR1 was expressed consistently across this broad sampling of WT and most robustly in the primitive blastema. Notably, FXR1 labeled a specific self-renewing progenitor population of the fetal kidney.     

严重烫伤后高迁移率族蛋白-1基因表达的改变及意义

目的　观察烫伤后肝、肺组织高迁移率族蛋白-1（HMG-1）mRNA表达的变化规律及其与器官功能损害的关系。　方法　采用大鼠35%Ⅲ度烫伤模型,动物随机分为正常对照组、烫伤组和重组杀菌/通透性增加蛋白(rBPI21)治疗组,留取组织和血标本分别检测组织内毒素含量、HMG-1mRNA表达及器官功能指标。　结果　严重烫伤后早期肝、肺组织HMG-1基因表达改变不明显（正常对照组肝组织0.202±0.097;肺组织0.263±0.091）,伤后24h则明显增多(肝组织0.487±0.189;肺组织0.513±0.069,P＜0.05,0.01),且一直持续至伤后72h(肝组织0.687±0.142;肺组织0.520±0.076,P＜0.01)。给予rBPI21治疗可有效抑制肝和肺中内毒素水平的升高,并显著抑制肝、肺组织HMG-1mRNA水平(P＜0.01)。相关分析结果显示,肝组织HMG-1mRNA表达与血清谷丙转氨酶、谷草转氨酶水平、肺组织HMG-1mRNA表达与髓过氧化物酶活性呈显著正相关（P＜0.05,0.01）。　结论　严重烫伤后肝、肺组织HMG-1表达显著增多,且持续时间较长,局部组织HMG-1诱生与烧伤后内毒素介导的重要器官功能损害关系密切。     

肠缺血-再灌注大鼠不同组织ICAM-1表达的规律

目的:了解肠缺血-再灌流过程中不同脏器 ICAM-1表达的时间、空间规律。方法:用 RT-PCR 和免疫组化的方法检测肠缺血-再灌流大鼠肠、肝和肺组织中 ICAM-1mRNA 和蛋白表达的变化。结果:肠组织中的ICAM-1 mRNA 和蛋白在肠缺血期及再灌流1h 表达增加,肝脏和肺脏毛细血管内皮上的 ICAM-1表达增加在再灌流后2h 和6h,晚于肠组织,并且与组织中性白细胞的聚集增加一致。结论:肠缺血-再灌流大鼠不同组织的ICAM-1表达的变化呈现序贯性。     

大鼠脊神经前根切断后脊髓内Slit1表达的变化

目的 :探讨脊神经前根损伤后,大鼠脊髓组织中Slit1的表达变化,为进一步研究Slit1在神经再生中的作用提供依据。方法:应用2月龄SD大鼠共100只,体重250±20g,其中80只SD大鼠实施左侧L5及L6脊神经前根切断,分别在伤后1d、3d、7d、14d时处死(每个时间点20只),取L5~L6脊髓节段,标记左右;左侧半L5~L6脊髓节段为实验组;右侧半为自身对照组。假手术大鼠(20只)实施麻醉及暴露L5及L6脊神经手术,未行L5及L6脊神经前根切断术,取其L5~L6脊髓节段为空白对照组。采用免疫组化、Western blotting及RT-PCR法检测大鼠脊髓组织中Slit1的变化。结果:脊神经前根切断后1d、3d、7d、14d,实验组中Slit1阳性细胞数分别为5.78±1.53、15.85±2.65、23.93±1.53、8.78±1.78;自身对照组中分别为2.31±1.63、2.57±1.89、3.20±2.16、3.02±2.15。各时间点实验组中Slit1阳性细胞数显著高于空白对照组(2.89±1.26)及自身对照组(P0.05)。Western blotting示脊神经前根切断后1d、3d、7d、14d,实验组Slit1相对表达量分别为0.326±0.09、0.448±0.05、0.638±0.07、0.304±0.07;自身对照组分别为0.038±0.02、0.038±0.01、0.035±0.02、0.046±0.03。各时间点实验组Slit1相对表达量与空白对照组(0.038±0.03)及自身对照组比较,差异有显著性(P0.05)。RT-PCR示脊神经前根切断后1d、3d、7d、14d,实验组Slit1 m RNA相对量分别为0.380±0.03、0.425±0.04、0.768±0.05、0.605±0.04;自身对照组分别为0.210±0.04、0.265±0.03、0.292±0.02、0.261±0.02。各时间点实验组Slit1 m RNA相对量与空白对照组(0.231±0.03)及自身对照组比较,差异有显著性(P0.05)。结论 :脊神经前根切断后,同侧脊髓灰质前角内Slit1的表达显著增加。     

Changes in cardiac gene expression after pig-to-primate orthotopic xenotransplantation

Byrne GW, Du Z, Sun Z, Asmann YW, McGregor CGA. Changes in cardiac gene expression after pig‐to‐primate orthotopic xenotransplantation. Xenotransplantation 2011; 18: 14–27. © 2011 John Wiley & Sons A/S. Abstract: Background: Gene profiling methods have been widely useful for delineating changes in gene expression as an approach for gaining insight into the mechanism of rejection or disease pathology. Herein, we use gene profiling to compare changes in gene expression associated with different orthotopic cardiac xenotransplantation (OCXTx) outcomes and to identify potential effects of OCXTx on cardiac physiology. Methods: We used the Affymetrix GeneChip Porcine Genomic Array to characterize three types of orthotopic cardiac xenograft outcomes: 1) rejected hearts that underwent delayed xenograft rejection (DXR); 2) survivor hearts in which the xenograft was not rejected and recipient death was due to model complications; and 3) hearts which failed to provide sufficient circulatory support within the first 48 h of transplant, termed “perioperative cardiac xenograft dysfunction” (PCXD). Gene expression in each group was compared to control, not transplanted pig hearts, and changes in gene expression > 3 standard deviations (±3SD) from the control samples were analyzed. A bioinformatics analysis was used to identify enrichments in genes involved in Kyoto Encyclopedia of Genes and Genomes pathways and gene ontogeny molecular functions. Changes in gene expression were confirmed by quantitative RT‐PCR. Results: The ±3SD data set contained 260 probes, which minimally exhibited a 3.5‐fold change in gene expression compared to control pig hearts. Hierarchical cluster analysis segregated rejected, survivor and PCXD samples, indicating a unique change in gene expression for each group. All transplant outcomes shared a set of 21 probes with similarly altered expression, which were indicative of ongoing myocardial inflammation and injury. Some outcome‐specific changes in gene expression were identified. Bioinformatics analysis detected an enrichment of genes involved in protein, carbohydrate and branched amino acid metabolism, extracellular matrix–receptor interactions, focal adhesion, and cell communication. Conclusions: This is the first genome wide assessment of changes in cardiac gene expression after OCXTx. Hierarchical cluster analysis indicates a unique gene profile for each transplant outcome but additional samples will be required to define the unique classifier probe sets. Quantitative RT‐PCR confirmed that all transplants exhibited strong evidence of ongoing inflammation and myocardial injury consistent with the effects of cytokines and vascular antibody‐mediated inflammation. This was also consistent with bioinformatic analysis suggesting ongoing tissue repair in survivor and PCXD samples. Bioinformatics analysis suggests for the first time that xenotransplantation may affect cardiac metabolism in survivor and rejected samples. This study highlights the potential utility of molecular analysis to monitor xenograft function, to identify new molecular markers and to understand processes, which may contribute to DXR.     

大鼠肺移植术后早期肺组织ICAM-1表达的变化

目的　研究大鼠肺移植术后早期肺组织细胞间粘附分子 1(ICAM 1)表达的变化。方法　用免疫组织化学 (SABC)法及设置内参照的半定量RT PCR方法 ,对移植肺获再灌注及通气 4h后肺组织的ICAM 1蛋白及其mRNA表达水平进行观察。实验组 (n =6 )肺经低钾右旋糖酐液 4℃保存12h后 ,行同种异体左肺原位移植。对照组 (n =6 )充分游离左侧肺动、静脉及支气管。免疫组织化学检测结果按阴性 (- )、可疑 (± )、弱阳性 ( )、阳性 ( )、强阳性 ( )进行记录。PCR产物电泳后扫描记录条带密度。结果　移植肺肺泡上皮及肺血管内皮细胞免疫着色显著较对照组深 (P<0 .0 1)。其ICAM 1mRNA与β actinmRNART PCR扩增产物相对密度值亦显著高于对照组 ,分别为 0 .873± 0 .0 44和 0 .44 2± 0 .0 37,P <0 .0 1。结论　肺移植术后早期肺组织ICAM 1表达上调 ,这种上调与ICAM 1mRNA增强有关。     

Chemotherapy and P-glycoprotein expression in chondrosarcoma

Chondrosarcomas are alleged to be resistant to chemotherapy. A retrospective review of our experience primarily with dedifferentiated chondrosarcomas treated with chemotherapy was performed to re-evaluate the efficacy of chemotherapy for this tumor. There were 18 patients: 14 stage IIB and four stage III. Seventeen patients had dedifferentiated chondrosarcoma. The median age at diagnosis was 57 years. Fourteen of the patients underwent wide excision of the tumor, two underwent amputation, and two had no surgery. The femur and the pelvis were the most common locations of the primary tumor. Chemotherapy for 11 of the patients consisted of cisplatin and doxorubicin. Survival was analyzed with the Kaplan-Meier method; the median survival was 12 months. The hypothesis that chondrosarcomas express P-glycoprotein was tested. Expression of P-glycoprotein was evaluated by immunostaining with use of the C494 and C219 antibodies on 41 benign and malignant cartilage tumors, six of which were from the patients in the chemotherapy group. Immunostaining revealed that 37 of 41 cartilage tumors expressed P-glycoprotein. The rate of survival of patients with high-grade chondrosarcoma treated with chemotherapy is poor. P-glycoprotein expression is common in benign and malignant cartilage lesions. The lack of response to chemotherapy may be related to the expression of P-glycoprotein.     

Changes of occludin expression in intestinal mucosa after burn in rats

In severely burned rats, hyperemia, edema and other pathological injuries occur in the intestinal mucosa. Ultramicroscopically, the microvilli, tight junction and organelles are disrupted. Occludin is a functional component of tight junctions. The purpose of the present study is to investigate changes of occludin expression, and to further elucidate the relationship between occludin expression and ultrastructure damage. The fluorescence intensity of occludin was detected in intestinal wall by the method of immunofluorescence histochemistry and confocal laser scanning microscopy (CLSM). Expression of occludin and its mRNA were determined by western blotting and RT-PCR, respectively. Changes of intestinal mucosa ultrastructure were observed by TEM. The results showed that fluorescence intensity of occludin at 3PBH was enhanced, higher than that of the control group, being 80.77+/-8.38 and 72.86+/-4.74, respectively, and reached a peak at 12PBH (116.14+/-6.89). The expression levels of occludin at 3PBH and 6PBH were 1.21+/-0.02 and 1.53+/-0.14 times that of the control group, respectively, and there were significant differences (P<0.01) between 3PBH group and 6PBH group and control group. The levels of occludin mRNA were also enhanced. At 12PBH, the level reached a peak (P<0.01), being 2.00+/-0.24 times that of the control group. Coincidently, the structure of the tight junction between epithelial cells was disrupted on a large scale under TEM. We speculate that up-regulation of epithelial occludin may play a role in enhancing paracellular permeability and be related to the damage to the tight junction.     

Changes in expression of amyloid precursor protein and interleukin-1beta after experimental traumatic brain injury in rats

There is increasing evidence linking neurodegenerative mechanisms in Alzheimer's disease (AD) and traumatic brain injury (TBI), including increased production of amyloid precursor protein (APP), and amyloid-beta (Abeta) peptide. In vitro data indicate that expression of APP may be regulated in part by the inflammatory cytokine IL-1beta. To further investigate the mechanisms involved, we measured APP and IL-1beta protein levels and examined immunohistochemical localization of APP in brain tissue from rats subjected to controlled cortical impact (CCI) injury. Animals were examined at time intervals ranging from 3 h to 4 weeks after TBI. The 24-h time point revealed a dramatic increase in APP immunoreactivity, detected with both N- and C-terminal antibodies, in the hippocampus and cortex ipsilateral to injury. This finding was sustained up to 3 days post-injury. At these early time points, APP increase was particularly robust in the white matter axonal tracts. By 14 days after injury, APP immunoreactivity was not significantly different from sham controls in cortex, but remained slightly elevated in hippocampus. Western blot data corroborated early increases in hippocampal and cortical APP in injured versus control animals. Despite profound APP changes, no Abeta deposits were observed at any time after injury. Hippocampal and cortical IL-1beta increases were even more robust, with IL-1beta levels peaking by 6 h post-injury and returning to baseline by 24-72 h. Our results demonstrate that both APP and IL-1beta are rapidly elevated after injury. Because of the rapidity in the IL-1beta peak increase, it may serve a role in regulation of APP expression after TBI.     

周围神经挤压伤后纤溶成分表达的变化

目的 观察大鼠坐骨神经挤压伤后纤溶成分中组织型纤溶酶原激活剂（ｔ－ＰＡ）和纤溶酶原激活剂抑制因子１（ＰＡＩ－１）蛋白表达的变化,了解ｔ－ＰＡ、ＰＡＩ－１在周围神地操作后修复过程中的作用。方法 取２１只Ｗｉｓｔａｒ大鼠,分别造成坐骨神经挤压伤模型。在术后６ｈ、１ｄ、３ｄ、７ｄ、１４和２１ｄ分别取材进行组织学、免疫组化观察。结果 术后３ｄ时神经断端回缩球处的ｔ－ＰＡ抗原呈阳性反应。７ｄ时损伤进行组织学     

Rhabdomyosarcoma in adult prostate

A case of rhabdomyosarcoma of the adult prostate is reported. Metastatic disease in both lung fields was treated by triple-drug therapy. This regimen has been maintained to date with resolution of pulmonary nodules.     

中枢和外周神经损伤后胞体区看家基因表达的变化

目的观察并比较几种常用的看家基因,包括β-actin,GAPDH,18s RNA,cyclophilin A等在中枢和外周神经损伤后胞体分布区神经组织中表达的变化.方法Wistar雄性大鼠随机分为右侧坐骨神经夹伤,右侧坐骨神经结扎和胸段脊髓半横断三组.手术后7d,用相对定量RT-PCR的方法检测几种看家基因在受损神经元胞体分布区神经组织中的表达水平.结果GAPDH和Cyclophilin A的表达在不同模型中的变化幅度较小,损伤与对照侧比较差异均无显著性.β-actin和18sRNA的表达在不同模型中变化不同,在脊髓损伤和坐骨神经结扎后表达的变化有统计学意义.结论本研究结果为研究神经系统损伤后相关基因表达变化时内参照基因的选择提供参考,同时也为进一步认识看家基因更加广泛的功能提供依据.     

Testicular Function after Combined Chemotherapy for Metastatic Testicular Cancer

In 10 patients cured for metastatic testicular cancer by combination chemotherapy serum hormone levels and serum agglutinating antibodies were analysed 12 to 35 months after discontinuation of the treatment. Together with these examinations sperm analysis was done. All patients had increased levels of follicle stimulating hormone (FSH). Serum testosterone was usually in the lower part of the normal range (below 20 nmol/***l). In eight patients the serum agglutinating antibodies were normal, while two patients had increased levels. In all patients azoospermia was observed, indicating long-lasting infertility in patients testicular cancer treated by combination chemotherapy. The possible importance of cryopreservation prior to start of chemotherapy is discussed.     

Auricular Rhabdomyosarcoma in a Rat

An ear (auricular) neoplasm from a 1‐year‐old male rat was removed surgically and examined histologically. Macroscopically, the neoplasm was firm, white and measured (0.5 × 0.5 cm). Microscopically, the neoplasm was expansile, non‐encapsulated, and composed of large, pleomorphic, polygonal to spindle‐shaped cells containing multiple nuclei. Using immunohistochemical and chemical stains, the neoplastic cells were positive for vimentin, myoglobin, phosphotungstic acid haematoxylin and desmin, but had no immunoreactivity for cytokeratin or α‐smooth muscle actin. On the basis of histopathological, immunohistochemical and histochemical stains, a diagnosis of auricular rhabdomyosarcoma was made. Although reported infrequently in human, this is, to the author's knowledge, the first report that describes the detailed gross, histopathological, histochemical and immunohistochemical findings of auricular rhabdomyosarcoma in a rat.     

Rhabdomyosarcoma in an adult hand

Abstract

Rhabdomyosarcoma (RMS) is a malign tumour which arises from cells committed to a skeletal muscle lineage. It constitutes 4%–8% of all childhood malignancies but is rare in adults. The rare pleomorphic subtype occurs almost exclusively in adults and most often involves the extremities. RMS of the hand or foot comprise a minority of extremity cases. An adult patient with rhabdomyosarcoma in the hand, which is very rare, is presented in this article. General characteristics of the tumour and the treatment strategies are discussed.